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Biomedical subjects

M Takumida

Publications and source records attributed to M Takumida.

At least 55 records · Page 3Linked to original sources

High-resolution scanning electron microscopy of the murine olfactory receptor.

The anatomy of the murine olfactory receptor was analyzed using high-resolution scanning electron microscopy with the aldehyde-osmium-dimethyl sulfoxide-osmium method. A good three-dimensional image of the receptor cell architecture was obtained. Within the olfactory vesicle, a complex tubular network was found that seemed to be physically associated with adjacent organelles, the basal aspect of cilia, and the lateral plasmalemma. It is assumed that these structures may be of significance during the olfactory transduction process.

Animals↗

Functional aspects of the vestibular dark cells in the guinea pig: morphological investigation using ruthenium red staining technique.

The functional significance of the vestibular dark cells in the guinea pig was investigated using ruthenium red staining technique. The apical cell surface of the dark cell was covered by the fuzzy layer of the glycocalyx, which could be responsible for water and ion transport across the apical plasma membrane. It has been suggested that the otoconia have been at first demineralized by an unknown mechanism when they are attached to the dark cell surface, and the mucopolysaccharide and mucoproteins in the otoconial matrix are then absorbed by active pinocytosis in the dark cells. These functions may be closely related to the possible regulation of the calcium ion in the vestibular endolymph.

Animals↗

Glycocalyx and ciliary interconnections of the human vestibular end organs: an investigation by scanning electron microscopy.

The ultrastructure of glycocalyx and ciliary interconnections of the human vestibular end organs was investigated in a three-dimensional way by using scanning electron microscopy. The plasma membrane showed a rather rough appearance, which was most prominent in the ciliary membrane of sensory cells. Ciliary interconnections emerged from the plasma membrane and tightly connected neighbouring cilia. The tip links stretched from the tips of the stereocilia to their taller neighbours. These tip links were arranged in the direction of the kinocilium. These findings indicate that the ciliary interconnections may keep the cilia arranged in a bundle and the tip links may be involved in the sensory cell transduction system.

Cilia↗

Effect of dosing schedule on aminoglycoside ototoxicity: comparative cochlear ototoxicity of amikacin and isepamicin.

The effect of the dosing schedule on aminoglycoside ototoxicity was investigated in the guinea pig. The animals were given amikacin or isepamicin either by once-daily intramuscular injection of 200 mg/kg or by twice-daily injections of 100 mg/kg for 28 days. The once-daily treatment induced a lesser degree of ototoxicity than the twice-daily injections, which may indicate that the once-daily treatment therapy has a potential value in the clinical use of aminoglycoside antibiotics. Isepamicin sulfate is a new aminoglycoside antibiotic currently undergoing clinical investigation. This study revealed that isepamicin also has ototoxic properties as has been observed in all other aminoglycosides. The degree of ototoxicity, however, was markedly less than that of amikacin.

Amikacin↗

The endolymphatic sac and inner ear homeostasis. I: Effect of glycerol on the endolymphatic sac with or without colchicine pretreatment.

The combined effects of glycerol and colchicine on the endolymphatic sac were investigated in mice. Glycerol induced signs of secretion from the epithelium with formation of secretory granules in the light epithelial cells. Other characteristics of the epithelial lining were also changed resulting in an increased widening of the lateral intercellular spaces, a partial collapse of the lumen and with a deposition of a stainable substance within the lumen. This reaction lasted from 30 min to 24 h following the injection. Pretreatment with colchicine was found to decrease or inhibit the glycerol-induced secretion of macromolecules into the sac. The lumen collapsed but frequently there was no presence of stainable substance. Animals treated with both glycerol and colchicine showed marked signs of inner ear malfunction which could indicate that the secretory activity in the sac might be closely related to the regulation of inner ear fluid homeostasis and that functional disturbances in this system may lead to disorders of inner ear function.

Animals↗

The endolymphatic sac and inner ear homeostasis. II: Effect of glycerol on the sensory end organs with or without colchicine pretreatment.

The effects of glycerol and colchicine on the sensory end organs of the inner ear were investigated in mice. Glycerol alone induced a widening of the intercellular spaces lining vestibular dark and transitional cells as well as the marginal cells of the stria vascularis. This was noted within 30 min after the injection of glycerol and was normalized again within 4 h after the injection. Colchicine induced some morphological changes in the inner ear sensory cells, such as dissociation of Golgi complexes etc. These isolated glycerol or colchicine injections did not cause any signs of inner ear functional impairment. Treatment with glycerol following pretreatment with colchicine, however, induced marked inner ear dysfunction with impaired sense of balance and audition. The inner ear morphology revealed a combination of changes as compared with what was observed after isolated treatment with glycerol or colchicine i.e. edema of the stria vascularis, and vestibular dark and transitional cells as well as dissociation of Golgi complexes in the sensory cells. The cochlea showed moderate endolymphatic hydrops. These findings indicate that colchicine affects the inner ear fluid regulating mechanisms which may lead to severe functional derangement after additional glycerol treatment. It is conceivable that the present experiment may serve as a useful model for further studies on inner ear changes related to endolymphatic hydrops and Ménière's disease.

Animals↗

Initial changes in the sensory hair-cell membrane following aminoglycoside administration in a guinea pig model.

This study demonstrates the initial changes affecting the sensory hair-cell plasma membranes in the vestibular end organs of gentamicin-treated guinea pigs by using a ruthenium red staining technique. First, 0.1 ml of a solution containing 5 mg gentamicin sulfate was injected into the middle ear. After 7 days, the sensory hair cell cilia were observed to be degenerating. The various stages of this degeneration process were classified into two types: the decrease in glycocalyx was designated type I fusion, while type II fusion was characterized by a bleb formation of the plasma membrane of the sensory hair cells, followed by a decrease in glycocalyx. The latter mechanism allowed plasma membrane contact, with subsequent fusion of the plasma membrane of neighboring sensory hair-cell cilia. The material also illustrates the degeneration of ciliary actin filaments. These findings suggest that the aminoglycoside affects both the glycocalyx and the plasma membrane, and that the decrease in glycocalyx may be the first sign of sensory hair-cell fusion.

Animals↗

Ultrastructural localization of carbonic anhydrase in the vestibular end organs of the guinea pig.

Carbonic anhydrase activity was demonstrated cytochemically on an ultrastructural level in the vestibular end organs of the guinea pig. Reaction product was found in the dark cells, transitional cells, cells of the planum semilunatum and supporting cells. In the dark cells, reaction product was observed in the cytoplasm as well as in the basal infoldings. Reaction product was also observed in the basal infoldings of the transitional cells and the cells of the planum semilunatum. The globular structures inside the supporting cells, transitional cells and the cells of the planum semilunatum were also surrounded by the reaction product. These findings suggest that carbonic anhydrase may have different functions, such as water and ion transport, respiration, nutrition and calcium carbonate deposition in the vestibular end organs.

Animals↗

Lectin detection of carbohydrates in the endolymphatic sac.

The carbohydrate contents of the guinea pig endolymphatic sac were investigated by the use of lectins. The lumen of the endolymphatic sac was filled with stainable precipitate containing N-acetyl glucosamine, mannose, glucose, galactose and fucose. N-Acetyl galactosamine was also detected but in minute amounts. This composition corresponded to other areas in the inner ear, such as the cupula, the otolithic membrane and the tectorial membrane. The function of these carbohydrates may play an important role in preventing the lumen of the endolymphatic sac from collapsing as well as in regulating transepithelial fluid transport.

Acetylgalactosamine↗

The effect of gentamicin on the glycocalyx and the ciliary interconnections in vestibular sensory cells: a high resolution scanning electron microscopic investigation.

Changes in glycocalyx structure and ciliary interconnections of the vestibular sensory cells are demonstrated after gentamicin administration. A special high resolution scanning electron microscope and a tannic acid-osmium staining technique giving an almost three dimensional view were used to achieve this purpose. Guinea pigs were injected with a single dose of 5 mg of gentamicin directly into the middle ear. Seven days after the injection, it was possible to observe the degenerative process of the glycocalyx and the ciliary interconnections. The first detectable change was a disarrangement of the cilia with a loosening of the interconnections. The ciliary membrane presented with an irregular appearance. The tip links connecting the tips of the stereocilia to their neighbours were also affected showing elongation or even disappearance. In the later stages of the degeneration process, the sensory hairs presented with different degrees of fusion whereafter they finally disappeared totally. These findings suggest that the glycocalyx acts to maintain a normal stability and shape of the ciliary membrane and to keep the regular distance between cilia in order to maintain the arrangement of the whole ciliary bundle. Gentamicin probably affects the glycocalyx and the ciliary interconnections resulting in a disarrangement, detachment and fusion of cilia. The tip links, which are suggested to be involved in sensory cell transduction, seem to be also affected by gentamicin.

Animals↗

Epithelial cell surface morphology in the endolymphatic sac: a scanning electron microscopic study in the mouse.

The apical, lateral and basal surface structures of the epithelial cells in the murine endolymphatic sac were studied using the freeze-cracking technique and scanning electron microscopy. In this way, it was possible to visualize the luminal surface and the interior of the cell simultaneously. The light epithelial cells, with their smooth rounded nuclei and many mitochondria, had numerous microvilli on their apical cell surface, whereas the dark epithelial cells, with indented nuclei, had only a few such microvilli. The lateral surfaces of these cells were flat, with few projections facing a dilated lateral intercellular space.

Animals↗

Carbohydrates of the guinea pig vestibular supporting cells.

Ultrastructural localization of glycoconjugates with special reference to the synthesizing process was studied in the guinea pig vestibular supporting cells using the tannic acid and ruthenium red staining technique. The extracellular glycoconjugate (glycocalyx) was well stained by the ruthenium red which may be synthesized by GERL complex. The supporting cell has a numerous number of granules which also contain tannic acid positive glycoconjugate. These granules were closely related to the Golgi complex as well as the apical cell surface. It may be indicated that the glycoconjugate of the supporting cells is glycosylated at the Golgi complex, stored in the granules, transported, secreted, and form otoconial membrane and the cupula.

Acoustic Maculae↗

Functional aspects of carbohydrate complex of the statoconial membrane of the guinea pig utricular macula.

A functional aspect of the carbohydrate complex of the statoconial membrane of the guinea pig utricular macula was observed by the use of the lectins as well as the ruthenium red staining techniques. The ruthenium red staining technique visualized the whole structure of the statoconial membrane as well as the contact zone between the sensory epithelium and the statoconial membrane. The statoconial membrane is composed of the otoconial layer, gelatinous layer, and subcupular meshwork. The otoconial layer was interconnected with sensory epithelium by the gelatinous layer and subcupular meshwork which formed honeycomb structure and housed the sensory hair bundle. This may allow the free movement of the sensory hair bundle. The gelatinous layer and the subcupular meshwork contain N-acetylglucosamine, mannose, and galactose which form acidic mucopolysaccharide and glycoprotein. It has been indicated that these carbohydrate complex may play an important role for the mechanical coupling not only between the statoconial membrane and the sensory epithelium but also between the sensory hair bundle and the statoconial membrane.

Acoustic Maculae↗

Effect of gentamicin on the carbohydrates of the vestibular end organs: an investigation by the use of FITC-lectins.

The effect of gentamicin on the glycoconjugates in the vestibular end organs is demonstrated using FITC-lectins. Five milligrams of gentamicin dissolved in 0.1 ml. saline was injected in a single dose into the middle ear of adult guinea pigs. Seven days after the injection, the fluorescent reactivity of Wheat germ agglutinin (WGA), Abrus precatorius agglutinin (APA), Concanavaline A (Con A), and Latyrus odoratus agglutinin (LOA) was decreased in the apical epithelial cell surface as well as in the gelatinous layer of the otolithic membrane of the maculae. In the cupula of the crista ampullaris, the reactivity for WGA and Con A was reduced. The otoconia, however, showed no detectable changes. This indicates that gentamicin may induce an altered carbohydrate metabolism resulting in a decrease of N-acetyl-glucosamine, mannose, galactose and glucose in the glycocalyx lining the epithelial cells as well as in the gelatinous layer of the otolithic membrane or cupula in the vestibular end organs.

Acoustic Maculae↗

Effect of acoustic overstimulation on the glycocalyx and the ciliary interconnections in the organ of Corti: high resolution scanning electron microscopic investigation.

Changes in ciliary interconnections in the organ of Corti are described after acoustic overstimulation using a special high resolution scanning electron microscope and tannic acid-osmium staining technique, giving an almost three dimensional view. Guinea pigs were exposed to a 3.85 kHz pure tone at an intensity of 120 dB for 22.5 minutes. The first detectable change was a disarrangement of the cilia with a loosening of the interconnections. The ciliary plasma membrane presented with an abnormally smooth appearance. The tip links connecting the tips of the stereocilia to their taller neighbours were also affected showing elongation or even disappearance. The fine granules which cover the tips of the tallest stereocilia of the outer hair cells were decreased. These findings suggest that acoustic overstimulation may affect the carbohydrate metabolism exceeding to degeneration of ciliary interconnections resulting in a disarrangement and detachment of cilia. The tip links, which may participate in sensory cell transduction, seem also to be affected by acoustic overstimulation.

Acoustic Stimulation↗

Effects of amiloride on the endolymphatic sac.

The effect of amiloride on the murine endolymphatic sac was investigated. The amiloride caused collapse of the lateral intercellular spaces in the endolymphatic sac epithelium and a subsequent mild endolymphatic hydrops. These changes indicated a decreased absorption of endolymph in the endolymphatic sac. Amiloride is known to inhibit the transcellular fluid transport without inducing any changes in the paracellular fluid transport. It is therefore suggested that amiloride specially inhibits the fluid and ion exchange in the apical portion of the epithelial cells resulting a decrease in transcellular fluid transport across the endolymphatic sac epithelium. The transcellular fluid transport seems to be one of the main mechanisms in the endolymphatic sac fluid exchange system.

Amiloride↗

Turnover of sulphur compounds in the endolymphatic sac: an autoradiographic study in the Mongolian gerbil.

Complex macromolecules are suggested to play an important role for the function of the endolymphatic sac (ES). As proposed in previous experimental studies, proteoglycans are supposed to be present in the ES. As they are composed of a proteic core linked to chains of glycosaminoglycans, chemical identification of the glycosaminoglycans is of particular importance, bearing in mind that all but hyaluronic acid contain sulphur. In order to follow the short-term turnover of sulphur in the ES, an autoradiographic study has been carried out in the Mongolian gerbil using 35S as tracer. Radioactive labelling of the gerbilline ES was controlled 15, 20, 30 and 60 min after intraperitoneal injection. While the first signs of the presence of radioactive sulphur were noticed after 20 min in the blood vessels and in the basal aspect of the ES epithelium, after 60 min it was possible to observe the presence of the tracer both within the epithelial cell layer as well as in the lumen of the ES. These findings are consistent with the presence of a fast turnover of sulphur molecules in the gerbilline ES.

Animals↗

Synthesis of glycocalyx and associated structures in vestibular sensory cells.

The ultrastructure of the glycocalyx with special reference to the synthesizing process was studied in the guinea pig vestibular sensory cells using the tannic acid staining technique. The glycocalyx emerged from the outer layer of the plasma membrane covering the entire length of the cilia. This glycocalyx also interconnected the ciliary structures tightly, such that a structural continuity was established between actin-membrane links and the glycocalyx. Interconnections between the actin filaments themselves were also noticed in the stereocilia as well as interconnections between individual actin filaments and the plasma membrane. These findings indicate that the glycocalyx and the ciliary interconnections may be closely related to the sensory hair transduction system. In the cellular cytoplasm, vesicles seemingly related to the synthesis of the glycocalyx were observed. These coated vesicles, which were synthesized by the Golgi complex and endoplasmic reticulum, interacted with the plasma membrane forming a coated pit. The lysosomal-like bodies also observed in the cell were closely related to the glycocalyx as well. Thus the glycocalyx seems to be synthesized by the endoplasmic reticulum and Golgi complexes and transferred through the coated vesicles or lysosomal-like bodies to the apical plasma membrane.

Animals↗